Hoechst 33258 Counterstaining
- the Hoechst precipitates with PO4, the embryos must be therefore treated in PO4 free buffer.
- Hoechst stock concentration is 1 mg/ml => must be diluted to the right concentration in any PO4 free buffer (Example: 0.1M Tris, pH 7.5).
- Incubate for +/- 30 min.
- Rinse in the buffer that has been used to prepare the mounting solution (see mounting).
Mounting
- Mounting is usually made in 70% glycerol (70% glycerol + 30% buffer)